CN111040975A - Enzyme-producing bacillus halophilus and application thereof in preventing and controlling pathogenic bacteria in saline water aquaculture - Google Patents
Enzyme-producing bacillus halophilus and application thereof in preventing and controlling pathogenic bacteria in saline water aquaculture Download PDFInfo
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Abstract
The invention discloses an enzyme-producing bacillus halophilus and application thereof in preventing and treating pathogenic bacteria in saline water aquaculture. The bacillus was named: bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 with the deposition number of GDMCC NO: 60904. The bacillus amyloliquefaciens CYZF1-2 has good enzyme production capacity: the hydrolysis effect on protein, starch and fat is good; the strain grows best in a culture medium with a salt concentration of 4%, and the OD of a bacterial liquid after 48h of culture600A value close to 1.0; the strain CYZF1-2 has high-efficiency energy inhibition effect on common aquatic pathogenic bacteriaThe antibacterial substance is mainly protein polypeptide substance, is used for preventing and treating pathogenic bacteria, does not generate drug resistance and secondary pollution, and has great application potential in preventing and treating diseases of saline water aquaculture.
Description
Technical Field
The invention relates to the technical field of application of aquatic microorganisms, in particular to a bacillus halophilus and application thereof in preventing and controlling pathogenic bacteria in saline aquiculture.
Background
China is the only world with the mariculture yield exceeding the marine fishing amount, and according to reports of world fishery and aquaculture conditions issued by FAO, the mariculture yield of China accounts for more than 80% of the world mariculture yield throughout the year 2013. Seawater or saline water culture plays an important role in aquaculture in China. However, in recent years, the aquaculture industry is rapidly developed in a large-scale and intensive direction, and brings many environmental problems and ecological crisis while promoting the development of agricultural economy and improving the quality of life of people. With the expansion of the scale of seawater or saline water culture and the continuous deterioration of culture environment, the disease problem in aquaculture, especially bacterial diseases, frequently occur in a high-density culture system and have serious influence, and the disease problem is fulminant and popular, so the disease problem becomes an important factor for restricting the sustainable and healthy development of the global mariculture industry, and the sustainable and healthy development of the national aquaculture industry is greatly limited. Among them, vibriosis such as Vibrio harveyi (Vibrio harveyi), Aeromonas disease such as Aeromonas hydrophila (Aeromonas hydrophila), edwardsiellosis such as Edwardsiella tarda (edwards siella tarda), etc. in the aquaculture process are the most common pathogenic bacteria, often causing large-scale outbreak of diseases related to aquaculture animals. In addition, Staphylococcus aureus (Staphylococcus aureus), salmonella typhimurium (salmonella typhimurium), Escherichia coli (Escherichia coli), and the like are common pathogenic bacteria in the intestinal tract or attached to feed, and are easily overlooked, but have a great influence on the health of cultured animals. At present, antibiotic drugs are widely applied to the treatment of bacterial diseases as efficient antibacterial drugs, and have good treatment effects, and meanwhile, the normal flora of aquaculture water is damaged, so that the drug resistance of pathogenic strains is enhanced, drug-resistant genes are enriched in fish bodies, and the ecological environment and the human health are seriously threatened.
Microbial agents (probiotics) are gradually favored by the market due to their advantages of safety, no toxicity, no residue, no pollution and the like. The biological control is carried out by utilizing the antagonism among microorganisms, and the method becomes an important effective way for controlling bacterial pathogens of aquatic products. However, related researches are still very limited, and particularly, antagonistic probiotics for efficient salt-loving saline aquaculture are reported, so that strain resources are very deficient. Therefore, the strain which is salt-loving and has high-efficiency aquatic pathogenic bacterium inhibiting capability is separated and screened, and is applied to the saline water aquaculture, and the method has important practical significance for realizing the real antibiotic-free aquaculture in the aquaculture industry.
Disclosure of Invention
The first purpose of the invention is to provide a Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 which can produce enzyme and salt and has the capability of efficiently antagonizing common aquaculture pathogenic bacteria, and a good biological material is provided for preventing and treating the aquaculture pathogenic bacteria.
The second purpose of the invention is to provide the application of the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 in the control of pathogenic bacteria in aquaculture, in particular the application in the control of pathogenic bacteria in saline water aquaculture.
Preferably, the application of the composition in preparing preparations for preventing and controlling Edwardsiella tarda (Edwards siella tarda), Aeromonas hydrophila (Aeromonas hydrophylla), Vibrio harveyi (Vibrio harveyi), Staphylococcus aureus (Staphylococcus aureus) and Salmonella typhimurium (Salmonella typhimurium) in saline water aquaculture.
Further preferably, the bacillus CYZF1-2 is applied to aquaculture with the salinity of 0-8%.
The third purpose of the invention is to provide a pathogenic bacteria prevention and control preparation for saline water aquaculture, which contains bacillus CYZF1-2 as an active ingredient.
The pathogenic bacteria preventing and treating preparation for the saline water aquaculture is a preparation for preventing and treating Edwardsiella tarda, Aeromonas hydrophila, Vibrio harveyi, Staphylococcus aureus and Salmonella typhimurium in the saline water aquaculture.
The fourth purpose of the invention is to provide the application of the bacillus CYZF1-2 in preparing protease, amylase and/or lipase.
The fifth purpose of the invention is to provide the application of the bacillus CYZF1-2 in preparing the medicines for resisting Edwardsiella tarda, Aeromonas hydrophila, Vibrio harveyi, Staphylococcus aureus and Salmonella typhimurium.
The sixth purpose of the invention is to provide a medicine for resisting Edwardsiella tarda, Aeromonas hydrophila, Vibrio harveyi, Staphylococcus aureus or Salmonella typhimurium, which contains bacillus CYZF1-2 as an active ingredient.
The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 is obtained by secondary domestication, enrichment and separation of pig manure in 2019 in 3 months by an inventor, and has the physiological and biochemical characteristics that: gram-positive bacteria, wherein the cells are short rods with spores and 0.5-1.0 multiplied by 1.5-2.5 mu m in size (see figure 1); the colony is white, the surface is rough, and the edge is irregular. Can grow vigorously under the condition of salinity of 4 percent.
Extracting the genome DNA of the halophilic antibacterial bacillus CYZF1-2, amplifying the 16SrDNA gene by using a 27F/1492R primer, and sequencing to obtain a gene sequence shown in SEQ ID NO. 1. The sequence is subjected to homology comparison analysis on NCBI and EzBioCloud websites to construct a phylogenetic tree, and morphological observation is combined, so that the strain is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens).
Compared with the prior art, the invention has the following beneficial effects:
(1) the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 has stronger capability of producing protease, amylase and lipase.
(2) The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 is a halophilic strain, grows most vigorously under the environment with salinity of 4 percent, and is more suitable for growing in the environment with lower salinity (the salinity is less than or equal to 2 percent); the growth can be well performed under the condition that the salinity is 6 percent, and the growth is obviously inhibited under the condition that the salinity is 8 percent.
(3) The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 can generate antibacterial peptide substances and has good antibacterial effect on common aquaculture pathogenic bacteria.
In conclusion, the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 has strong growth adaptability in seawater or saline water, has good bacteriostatic effect on common aquaculture pathogenic bacteria, and has great application potential in saline water aquaculture.
The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1-2 is preserved in Guangdong province microbial culture collection center (GDMCC) in 2019, 12 and 25 months, and is addressed to No. 59 large yard of Zuixianrui No. 100 of Guangdong province, Guangzhou city, Vibrio Xiuqing, and the preservation number is GDMCC NO: 60904.
Drawings
FIG. 1 is a photograph of Bacillus amyloliquefaciens CYZF1-2 stained with crystal violet under an optical microscope (100X).
FIG. 2 is a bar chart of the enzyme production ability evaluation of Bacillus amyloliquefaciens CYZF1-2, wherein different substrates are hydrolyzed, and the enzyme production ability is stronger when the ratio of the diameter of a hydrolysis transparent ring to the diameter of a colony is larger.
FIG. 3 is a graph showing the effect of salt concentration conditions on the growth of Bacillus amyloliquefaciens CYZF 1-2.
FIG. 4 is a bar graph showing the inhibitory effect of Bacillus amyloliquefaciens CYZF1-2 on 5 pathogenic bacteria.
FIG. 5 is a photograph showing the inhibition zone of the supernatant after centrifugation of the Bacillus amyloliquefaciens CYZF1-2 fermentation broth on pathogenic bacteria strains GDMCC 1.781(Vibrio harveyi 1.781) and GDMCC 1.379(Edwards siella tarda 1.379), wherein a is the strain GDMCC 1.781, and b is the strain GDMCC 1.379.
FIG. 6 is a phylogenetic tree of Bacillus amyloliquefaciens CYZF1-2 and related strains; where the tree was built using the NJ method, only bootstrap coefficients > 70% are shown (1000 replicates).
Detailed Description
The following detailed description of the embodiments of the present invention will be described in detail with reference to the accompanying drawings and examples, which are provided for illustration of the present invention and are not intended to limit the scope of the present invention, and the parameters, proportions, etc. of the examples may be selected according to circumstances without substantially affecting the results. Unless otherwise specified, the methods described in the examples are all conventional methods, and the reagents used are all conventional reagents or reagents formulated in a conventional manner.
Example 1 enrichment culture, isolation, purification and preservation of Bacillus amyloliquefaciens CYZF1-2
(1) Enrichment culture
Enrichment culture medium: NH (NH)4Cl 0.4g/L、NaNO20.25 g/L、KH2PO42.0 g/L、MgSO4·7H2O 0.2g/L、Na2CO30.4 g/L, and the solvent is water; the preparation method comprises the following steps: mixing the above components, and sterilizing at 121 deg.C under high temperature and high pressure for 20 min. The mixture was dispensed into 250mL triangular flasks, each 90 mL.
Collecting fresh pig manure 10g, cold preserving at 4 deg.C, placing into conical flask containing the above culture medium, culturing at 30 deg.C and 180rpm for 48 hr, and performing enrichment culture. Then transferring the culture medium into a new conical flask filled with the enrichment culture medium for culturing for 48 hours at 30 ℃ and 180rpm, and carrying out secondary enrichment to obtain a secondary enrichment culture.
(2) Separation, purification and preservation of bacillus CYZF1-2
Isolation medium (nutrient broth medium): 5g of beef extract, 10g of peptone, 5g of sodium chloride and 15g of agar, adding water to a constant volume of 1000mL, adjusting the pH value to 7.4, and subpackaging into 250mL triangular bottles with each bottle being 100 mL; sterilizing at 121 deg.C under high temperature and high pressure for 20min, and pouring into flat plate.
Coating the secondary enrichment culture obtained in the step (1) on a flat plate containing a separation culture medium by adopting a dilution coating method; observing the colony morphology by naked eyes, wherein the colony is white, rough in surface and irregular in edge; meanwhile, the cells are observed to be short-rod-shaped, have spores, have the size of 0.5-1.0 multiplied by 1.5-2.5 mu m and are gram-positive bacteria under an oil microscope with the power of 100 times of that of the cells by crystal violet staining (figure 1), and the results are shown in figure 1; further streaking and purifying, and carrying out amplification culture on the purified strain and preserving the strain in a glycerol tube (-80 ℃) and a freeze-dried tube (4 ℃), thereby obtaining the strain CYZF 1-2.
Example 2 identification of 16S rDNA of Bacillus amyloliquefaciens CYZF1-2
Extracting genome DNA of a strain CYZF1-2, amplifying by using a bacterial 16S rDNA gene amplification universal primer 27F/1492R (5'-AGAGTTTGATCCTGGCTCAG-3' and 5'-TACGACTTAACCCCAATCGC-3') to obtain a PCR product, and sending the PCR product to Shanghai Meiji biological medicine science and technology limited company (Guangzhou division) for sequence sequencing, wherein the sequence length of the 16S rRNA obtained after sequencing is 1410bp, and the sequence is shown in SEQ ID NO. 1. The sequencing results were compared with the 16SrDNA sequences in NCBI and EzBioCloud databases for homology analysis, and then the strain CYZF1-2 and the similar strains were selected to use MEGA 6.0, Kimura2-parameter model, NJ algorithm to build phylogenetic tree (bootstrap repeats 1000 times), and the obtained results are shown in FIG. 6.
The analysis of the result shows that the similarity of the 16S rDNA gene sequence of the strain Brevibacterium frigoritolans and the 16S rDNA gene sequence of the strain Brevibacterium frigoritolans is the highest (99.48 percent).
In the ezbiocoud database, the 16S rRNA gene sequence of Bacillus (Bacillus sp.) CYZF1-2 (i.e. strain CYZF1-2) has the highest similarity (99.59%) to b.subtilis subsp.sternaris D7XPN1, but the coverage is only 86.2%, followed by b.siamensis KCTC 13613 (99.57%). According to the results of the evolutionary tree, the strain CYZF1-2 and Siamese bacillus B.siamensis KCTC 13613 (99.57%) and Bacillus amyloliquefaciens DSM 7 (99.36%) are polymerized on one branch, but the colony of the Siamese bacillus is observed in a morphological way to be round, flat in surface, slightly protruded on the edge, white, 1-3mm in diameter and obviously different from the strain CYZF 1-2. Based on the analysis of the results above, it was preliminarily identified that the strain CYZF1-2 isolated in example 1 of the present invention is Bacillus amyloliquefaciens. It was named: bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF 1-2. The strain is preserved in Guangdong province microbial culture collection center (GDMCC) in 2019, 12 months and 25 days, and the address: no. 59 building of No. 100 Dazhong Jie of Xieli Zhonglu, Guangdong province, with the preservation number GDMCC No. 60904.
Example 3 evaluation of the enzyme-producing ability of Bacillus CYZF1-2
Enzyme production capacity was assessed using the plate hydrolysis loop assay:
the culture medium and reagents mainly used are as follows:
protein medium: 40.0g of skimmed milk powder, 10.0g of soluble starch, 3.0g of yeast extract, 2.0g of potato powder and 15.0g of agar, and distilled water is added to the mixture until the volume is 1000mL and the pH value is 7.0-7.2. Sterilizing at 121 deg.C under high temperature and high pressure for 20min, and pouring into flat plate.
Starch culture medium: 5.0g of soluble starch, 5.0g of peptone, 5.0g of beef extract, 5.0g of sodium chloride and 15.0g of agar, and adding distilled water to a constant volume of 1000mL, wherein the pH value is 7.0-7.2. Sterilizing at 121 deg.C under high temperature and high pressure for 20min, and pouring into flat plate.
Fat medium (tributyrin): using nutrient broth as a basic culture medium, adding 2mL of fat source and 15.0g/L of agar into every 100mL of the basic culture medium, and adding distilled water to a constant volume of 1000mL and a pH value of 7.0-7.2; wherein the fat source is 2% polyvinyl alcohol solution and tributyrin at a ratio of 1:9 (v/v). Sterilizing at 121 deg.C under high temperature and high pressure for 20min, and pouring into flat plate.
The activated strain CYZF1-2 is respectively point-connected to a protein, starch and fat culture medium plate, and is placed in an incubator at 30 ℃ for culturing for 48 hours, and then the ratio of the hydrolyzed transparent ring diameter to the bacterial colony diameter is measured (iodine indicator with mass fraction of 0.1% is added to the starch culture medium plate), so that the capability of the strain for secreting and producing protease, amylase and lipase can be qualitatively and directly reflected.
The transparent circle diameters and colony diameter ratios of the strains CYZF1-2 hydrolyzed protein, starch and fat obtained by the detection method are respectively 1.41, 2.30 and 2.71, and the results are shown in figure 2. As shown in fig. 2, the strain CYZF1-2 showed strong ability to produce protease, amylase and lipase.
Example 4 salinity assessment of growth Adaptation of Bacillus amyloliquefaciens CYZF1-2
(1) Preparing nutrient broth culture media with different salinity gradients: the nutrient broth is used as a basic culture medium, 2g, 4g, 6g, 8g and 10g of sodium chloride are added into each 100mL of the basic culture medium to prepare culture media with the salinity of 2%, 4%, 6%, 8% and 10%, the pH is adjusted to be 7.2, and the culture media are sterilized at the high temperature and the high pressure of 121 ℃.
(2) Inoculating and culturing: inoculating strain CYZF1-2 into nutrient broth culture medium with different salinity gradient, shake culturing at 30 deg.C and 180rpm/min, sampling after culturing for 0, 8, 12, 24, and 48 hr, and detecting OD600The values, results are shown in FIG. 3.
As shown in figure 3, the strain CYZF1-2 has salt preference in a certain salinity range, grows best in a culture medium with 4% of salinity, and OD is obtained after 24h culture600The value already exceeds 0.8, OD after 48h of culture600A value close to 1.0; at lower salinity (such as 2%), the growth of thallus is limited, and the OD after 48h of culture600The value is close to 0.8, but when the salinity is very high (such as 8 percent), the growth of the thallus is obviously inhibited, and the OD is obtained after 48 hours of culture600The value was close to 0.6 and the stationary phase had been reached.
Example 5 determination of the bacteriostatic Effect of Bacillus amyloliquefaciens CYZF1-2
(1) Preparing test bacteria clear liquid: inoculating the activated strain CYZF1-2 into nutrient broth culture medium, and culturing in a shaker at 30 deg.C and 180r/min for 24 h; centrifuging the culture solution, collecting supernatant, dividing into two parts, and standing one part at 4 deg.C; heating the other part in autoclave at 121 deg.C for 10 min.
(2) Preparing a bacterium-containing plate: the cultured test strains (shown in Table 1 below) were subjected to OD adjustment using an ultraviolet spectrophotometer600Taking out proper amount of bacterial liquid, adding into corresponding culture medium (2216E culture medium for Vibrio harveyi and nutrient broth culture medium for others) melted at about 45 deg.C, shaking, pouring into flat plate, naturally solidifying, and perforating with hole puncherThe same dish was perforated uniformly (aperture 9 mm).
(3) Bacteriostasis test
And (3) sucking 100 mu L of the centrifuged supernatant bacterial liquid of the strain CYZF1-2, respectively adding the supernatant bacterial liquid into each plate containing the pathogenic bacteria to be tested, culturing the plates in an incubator at 37 ℃ or 30 ℃ for about 48 hours, observing the bacteriostasis effect (the result is shown in figure 5), and measuring the diameter of the bacteriostasis zone (the result is shown in figure 4).
TABLE 1 Bacillus amyloliquefaciens CYZF1-2 bacteriostatic effect test strains
(4) Analysis of antibacterial substance by Strain CYZF1-2
1) Stability to high temperature test
And (3) sucking 100 mu L of supernatant of the strain CYZF1-2 heated in an autoclave at 121 ℃ for 10min, adding the supernatant into a plate containing the strain GDMCC 1.1220, culturing the plate in an incubator at 30 ℃ for about 48h, observing the bacteriostasis effect, and measuring the diameter of a bacteriostasis ring, wherein the result shows that the bacteriostasis ring is still obvious, and the ratio of the diameter to the diameter before sterilization is slightly smaller (14.60 +/-0.21 mm), which indicates that the antibacterial substance of the strain CYZF1-2 has good thermal stability.
2) Stability test for proteases
1mL of 2 parts of bacterial clear liquid (supernatant of a strain CYZF1-2, which is not sterilized) filtered by a filter head with the aperture of 0.22 mu m is taken, 30 mu L of neutral protease and 5g/L of compound protease (protease K and papain in a mass ratio of 1:1) solution are respectively added into the 1mL of bacterial clear liquid, 30 mu L of normal saline is added into a control group, and the experimental group and the control group are respectively treated in water bath at 37 ℃ for 1 h. The antibacterial effect of the treated bacteria clear liquid is determined by the punching method. Compared with a control group (the diameter of the inhibition zone is 15.0 +/-0.08 mm), the bacteriostatic activity of the protease-treated bacteria clear liquid is obviously reduced, and the inhibition zone is not obvious (the diameters of the inhibition zones treated by neutral protease and compound protease are 10.34 +/-0.089 mm and 9.8 +/-0.17 mm respectively), which shows that the antagonistic substance is sensitive to the protease, so that the antagonistic substance is judged to be a protein substance.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Sequence listing
<110> Guangdong province institute for microbiology (Guangdong province center for microbiological analysis and detection)
<120> one strain of enzyme-producing bacillus halophilus and application thereof in preventing and controlling pathogenic bacteria in saline water aquaculture
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<170>SIPOSequenceListing 1.0
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<211>1410
<212>DNA
<213> Bacillus amyloliquefaciens CYZF1-2(Bacillus amyloliquefaciens CYZF1-2)
<400>1
aaaggttacc tcaccgactt cgggtgttac aaactctcgt ggtgtgacgg gcggtgtgta 60
caaggcccgg gaacgtattc accgcggcat gctgatccgc gattactagc gattccagct 120
tcacgcagtc gagttgcaga ctgcgatccg aactgagaac agatttgtgg gattggctta 180
acctcgcggt ttcgctgccc tttgttctgt ccattgtagc acgtgtgtag cccaggtcat 240
aaggggcatg atgatttgac gtcatcccca ccttcctccg gtttgtcacc ggcagtcacc 300
ttagagtgcc caaactgaat gcctggcaac taagattcaa agggttgcgc tcgttgcggg 360
gacttaaccc aacatctcac gacacgagct gacgacaacc atgcaccacc tgtcactctg 420
cccccgaagg ggacgtccta tctctaggat tgtcagagga tgtcaagacc tggtaaggtt 480
cttcgcgttg cttcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc 540
ctttgagttt cagtcttgcg accgtactcc ccaggcggag tgcttaatgc gttagctgca 600
gcactaaggg gcggaaaccc cctaacactt agcactcatc gtttacggcg tggactacca 660
gggtatctaa tcctgttcgc tccccacgct ttcgctcctc agcgtcagtt acagaccaga 720
gagtcgcctt cgccactggt gttcctccac atctctacgc atttcaccgc tacacgtgga 780
attccactct cctcttctgc actcaagttc cccagtttcc aatgaccctc cccggttgag 840
ccgggggctt tcacatcaga cttaagaaac cgcctgcgag ccctttacgc ccaataattc 900
cggacaacgc ttgccaccta cgtattaccg cggctgctgg cacgtagtta gccgtggctt 960
tctgggttag gtaccgtcaa ggtgccgccc ctatttgaac ggcacttgtt cttccctaac 1020
aacagagctt tacgatccga aaaccttcat cactcacgcg gcgttgctcc gtcagacttt 1080
cgtccattgc ggaagattcc ctactgctgc ctcccgtagg agtctgggcc gtgtctcagt 1140
cccagtgtgg ccgatcaccc tctcaggtcg gctacgcatc gtcgccttgg tgagccgtta 1200
cctcaccaac tagctaatgc gccgcgggtc catctgtaag tggtagccga agccaccttt 1260
tatgtctgaa ccatgcggtt cagacaacca tccggtatta gccccggttt cccggagtta 1320
tcccagtctt acaggcaggt tacccacgtg ttactcaccc gtccgccgct aacatcaggg 1380
agcaagctcc catctgtccg ctcgactgca 1410
Claims (9)
1. A strain of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens CYZF1-2) CYZF1-2 has a preservation number of: GDMCC NO: 60904.
2. The application of the bacillus CYZF1-2 in preparing the preparation for preventing and treating pathogenic bacteria in saline water aquaculture.
3. The use according to claim 2, for the control of Edwardsiella tarda (edwards siella tarda), Aeromonas hydrophila (Aeromonas hydrophila), vibrio harveyi (vibrio harveyi), Staphylococcus aureus (Staphylococcus aureus), salmonella typhimurium (salmonella typhimurium) in saltwater aquaculture.
4. The use of claim 2 or 3, wherein the bacillus CYZF1-2 is used in aquaculture with salinity of 0-8%.
5. A preparation for controlling pathogenic bacteria in saline water aquaculture, which comprises the bacillus CYZF1-2 of claim 1 as an active ingredient.
6. The saltwater aquaculture pathogenic bacteria control formulation of claim 5 wherein said saltwater aquaculture pathogenic bacteria control formulation is an Edwardsiella tarda, Aeromonas hydrophila, Vibrio harveyi, Staphylococcus aureus, Salmonella typhimurium.
7. Use of bacillus CYZF1-2 according to claim 1 for the preparation of proteases, amylases and/or lipases.
8. The use of the bacillus CYZF1-2 of claim 1 in the preparation of medicines for resisting Edwardsiella tarda, Aeromonas hydrophila, Vibrio harveyi, Staphylococcus aureus and Salmonella typhimurium.
9. A drug against Edwardsiella tarda, Aeromonas hydrophila, Vibrio harveyi, Staphylococcus aureus or Salmonella typhimurium, which comprises the Bacillus CYZF1-2 of claim 1 as an active ingredient.
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